Four strains (Y4, Y9, Y13, N6) producing ethyl carbamate degrading enzymes were isolated and purified from 36 strains of yeast using a selective culture medium. All the ITS sequences were Pichia kudriavzevii. Through single-factor tests and orthogonal tests, the optimization of the fermentation conditions for EC-producing hydrolase of Y13 was studied. The results showed that maltose was used as the carbon source, the concentration was 10 g/L, the yeast extract was used as the nitrogen source, the concentration was 30 g/L, the inoculation amount was 6 %, and the EC substrate concentration in the medium was 5.00%. The optimum fermentation conditions were as follows: liquid volume 20 g/L, initial pH 6, culture temperature 28 ℃, and culture time 120 h